Analytische Chemie
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Biomedical applications, including functional biomaterials, carbohydrate-arrays, and glycan-based biosensors.
The chemistry of glycan immobilization plays an essential role in the bioavailability and function of surface bound carbohydrate moieties. For biomedical applications the stability over time (shelf life) of glycan arrays is a crucial factor. Herein we report on approaches for surface and interface characterization relevant to the needs of production of glycan microarrays which were tested using model carbohydrate surfaces. For detailed characterization of glycan model surfaces we used a combination of X-ray photoelectron spectroscopy (XPS), near edge X-ray absorption fine structure spectroscopy (NEXAFS) and ToF SIMS which are complementary techniques of surface chemical analysis. Links to fluorescence spectroscopy often used for characterization in the microarray community were established as well. In detail, amine-reactive silicon oxide and glass surfaces were used for anchoring oligosaccharides with an amino linker. The amount of surface bound carbohydrates was estimated by X-ray photoelectron spectroscopy (XPS). Glycan immobilization was investigated using lectins, which are glycan-binding molecules. A shelf life study of model glycan microarrays on epoxy-coated glass surfaces was done over a period of 160 days under different storage conditions utilizing fluorescence, ToF-SIMS and XPS analysis. It was shown that glycan activity of the models used can be maintained at least for half a year of storage at 4 °C.
Covalent modification of surfaces with carbohydrates (glycans) is a prerequisite for a variety of glycomics-based biomedical applications, including functional biomaterials, glycan-arrays, and glycan-based biosensors. The chemistry of glycan immobilization plays an essential role in the bioavailability and function of surface bound carbohydrate moieties. For biomedical applications the stability over time (shelf life) of a glycan-array is a crucial factor. Basic requirements for the production of microarrays are first of all stable signals without any loss of quality. Therefore, the investigation of the shelf life for carbohydrate microarrays is an important part in the development of glycan-arrays.
Motivated by the need of reliable quality control for glycan microarrays, we developed reference arrays using fluorescent model glycans. Since the long term stability of glycan microarrays is a crucial factor for their clinical application the shelf life at different storage conditions of glycan microarrays was studied in detail using the two model glycan compounds.
Herein, we present a shelf life study of model glycan microarrays on epoxy modified glass surfaces over a period of 320 days. This was carried out using different analyzing techniques such as Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS), X-ray Photoelectron Spectroscopy (XPS) and Fluorescence Spectroscopy. To analyze and interpret the ToF-SIMS dataset the multivariate technique principal component analysis (PCA) was used. The dependence of the array´s shelf life upon storage conditions was specifically studied.